Keywords
immunohistochemistry
PRP
combined topical preparations
complex treatment
tissue regeneration
macrophages
inflammation
endothelial cells
microvessel density
How to Cite
Abstract
Aim. To conduct a comparative analysis of CD34, CD68, and CD163 expression in tissues of patients with chronic wounds before and after complex treatment with combined topical preparations and adjunctive PRP and to assess additional cellular and humoral indicators of reparative dynamics.
Materials and methods. A comparative morphological, immunohistochemical, and clinical-laboratory study of biopsy material and local wound microenvironment indicators was performed in 40 patients with chronic wounds. The control group included 20 patients who received standard treatment; in the main group, tissue samples were evaluated before and after complex treatment in which combined topical preparations formed the basis of local therapy and PRP was used as an adjunct. CD34, CD68, and CD163 expression was assessed in representative high-power fields at ×400 magnification. In addition, wound imprint cytology, nitroblue tetrazolium test results for neutrophils and monocytes, TGF factor, reactive oxygen species, fibronectin, calpain, and pro-inflammatory cytokines were evaluated on days 1, 7, and 14.
Results. After treatment, the proportion of CD34-positive cells increased from 6.92% to 26.77%, indicating activation of angiogenesis. At the same time, CD68-positive cells decreased from 67.83% to 27.05%, and CD163-positive cells decreased from 73% to 40.9%, reflecting reduced macrophage infiltration and remodeling of the tissue microenvironment. In the main group treated with combined topical preparations and adjunctive PRP, the neutrophil content in wound imprints decreased from 73.3±6.0% to 55.9±6.2%, monocyte content increased from 3.9±1.7% to 10.2±1.2%, TGF factor increased from 100.8±6.3 to 131.4±5.9 pg/ml, and fibronectin increased from 146.6±3.9 to 182.0±2.5 pg/ml, while reactive oxygen species, calpain, and pro-inflammatory cytokines decreased.
Conclusions. During complex treatment with combined topical preparations and adjunctive PRP, comprehensive assessment of CD34, CD68, and CD163 expression combined with PRP-associated cellular and humoral indicators characterizes morphological and laboratory signs of chronic wound repair. The obtained data indicate activation of angiogenesis, reduction of the inflammatory macrophage component, and formation of a more favorable wound microenvironment. Because the intervention was multicomponent, the findings characterize the treatment regimen as a whole and do not establish the isolated effect of PRP or either topical preparation.
References
Nussbaum, S. R., Carter, M. J., Fife, C. E., DaVanzo, J., Haught, R., Nusgart, M., et al. (2018). An economic evaluation of the impact, cost, and Medicare policy implications of chronic nonhealing wounds. Value Health, 21(1), 27–32. https://doi.org/10.1016/j.jval.2017.07.007
Arif, M., Khan, S. M., Gull, N., Tabish, T. A., Zia, S., Khan, R. U., et al. (2021). Polymer-based biomaterials for chronic wound management: promises and challenges. International Journal of Pharmaceutics, 598, 120270. https://doi.org/10.1016/j.ijpharm.2021.120270
Eming, S. A., Martin, P., & Tomic-Canic, M. (2014). Wound repair and regeneration: mechanisms, signaling, and translation. Science Translational Medicine, 6(265), 265sr6. https://doi.org/10.1126/scitranslmed.3009337
Peña, O. A., & Martin, P. (2024). Cellular and molecular mechanisms of skin wound healing. Nature Reviews Molecular Cell Biology, 25(8), 599–616. https://doi.org/10.1038/s41580-024-00715-1
De Palma, M., Biziato, D., & Petrova, T. V. (2017). Microenvironmental regulation of tumour angiogenesis. Nature Reviews Cancer, 17(8), 457–474. https://doi.org/10.1038/nrc.2017.51
Annese, T., Tamma, R., De Giorgis, M., & Ribatti, D. (2020). microRNAs biogenesis, functions and role in tumor angiogenesis. Frontiers in Oncology, 10, 581007. https://doi.org/10.3389/fonc.2020.581007
Siemerink, M. J., Klaassen, I., Vogels, I. M. C., Griffioen, A. W., Van Noorden, C. J. F., & Schlingemann, R. O. (2012). CD34 marks angiogenic tip cells in human vascular endothelial cell cultures. Angiogenesis, 15(1), 151–163. https://doi.org/10.1007/s10456-011-9251-z
Wynn, T. A., & Vannella, K. M. (2016). Macrophages in tissue repair, regeneration, and fibrosis. Immunity, 44(3), 450–462. https://doi.org/10.1016/j.immuni.2016.02.015
Willenborg, S., Sanin, D. E., Jais, A., Ding, X., Ulas, T., Nüchel, J., et al. (2021). Mitochondrial metabolism coordinates stage-specific repair processes in macrophages during wound healing. Cell Metabolism, 33(12), 2398–2414.e9. https://doi.org/10.1016/j.cmet.2021.10.004
Krzyszczyk, P., Schloss, R., Palmer, A., & Berthiaume, F. (2018). The role of macrophages in acute and chronic wound healing and interventions to promote pro-wound healing phenotypes. Frontiers in Physiology, 9, 419. https://doi.org/10.3389/fphys.2018.00419
Hesketh, M., Sahin, K. B., West, Z. E., & Murray, R. Z. (2017). Macrophage phenotypes regulate scar formation and chronic wound healing. International Journal of Molecular Sciences, 18(7), 1545. https://doi.org/10.3390/ijms18071545
Kong, Q., Gao, S., Li, P., Sun, H., Zhang, Z., Yu, X., et al. (2024). Calcitonin gene-related peptide-modulated macrophage phenotypic alteration regulates angiogenesis in early bone healing. International Immunopharmacology, 130, 111766. https://doi.org/10.1016/j.intimp.2024.111766
Okonkwo, U. A., & DiPietro, L. A. (2017). Diabetes and wound angiogenesis. International Journal of Molecular Sciences, 18(7), 1419. https://doi.org/10.3390/ijms18071419
Radu, P., Zurzu, M., Paic, V., Bratucu, M., Garofil, D., Tigora, A., et al. (2023). CD34: structure, functions and relationship with cancer stem cells. Medicina (Kaunas), 59(5), 938. https://doi.org/10.3390/medicina59050938
Kumar, V., Abbas, A. K., & Aster, J. C. (Eds.). (2018). Robbins Basic Pathology (10th ed.). Elsevier. https://shop.elsevier.com/books/robbins-basic-pathology/kumar/978-0-323-35317-5
Dabbs, D. J. (Ed.). (2021). Diagnostic Immunohistochemistry: Theranostic and Genomic Applications (6th ed.). Elsevier. https://shop.elsevier.com/books/diagnostic-immunohistochemistry/dabbs/978-0-323-72172-1
Swain, S. S., Karri, V. V. S. R., Uddin, M. E., Gowthamarajan, K., Bhaskaran, M., & Subramaniyan, V. (2025). Advances in diabetic wound healing: from pathophysiology to emerging therapies. Acta Diabetologica. https://doi.org/10.1007/s00592-025-02629-6
Griffioen, A. W., & Dudley, A. C. (2022). The rising impact of angiogenesis research. Angiogenesis, 25(4), 435–437. https://doi.org/10.1007/s10456-022-09849-2
Woo, K., González, C. V. S., Amdie, F. Z., & de Gouveia Santos, V. L. C. (2024). Exploring the effect of wound-related pain on psychological stress, inflammatory response, and wound healing. International Wound Journal, 21(7), e14942. https://doi.org/10.1111/iwj.14942
Gruber, E., Heyward, C., Cameron, J., & Leifer, C. (2018). Toll-like receptor signaling in macrophages is regulated by extracellular substrate stiffness and Rho-associated coiled-coil kinase (ROCK1/2). International Immunology, 30(6), 267–278. https://doi.org/10.1093/intimm/dxy027
Bankhead, P., Loughrey, M. B., Fernández, J. A., Dombrowski, Y., McArt, D. G., Dunne, P. D., et al. (2017). QuPath: open source software for digital pathology image analysis. Scientific Reports, 7, 16878. https://doi.org/10.1038/s41598-017-17204-5
Rodrigues, M., Kosaric, N., Bonham, C. A., & Gurtner, G. C. (2019). Wound healing: a cellular perspective. Physiological Reviews, 99(1), 665–706. https://doi.org/10.1152/physrev.00067.2017
Altman, D. G., & Bland, J. M. (1999). Statistics notes: variables and parameters. BMJ, 318, 1667. https://doi.org/10.1136/bmj.318.7199.1667
McHugh, M. L. (2013). The chi-square test of independence. Biochemia Medica (Zagreb), 23(2), 143–149. https://doi.org/10.11613/bm.2013.018
Qu, W., Wang, Z., Hunt, C., Morrow, A. S., Urtecho, M., Amin, M., et al. (2021). The effectiveness and safety of platelet-rich plasma for chronic wounds: a systematic review and meta-analysis. Mayo Clinic Proceedings, 96(9), 2407–2417. https://doi.org/10.1016/j.mayocp.2021.01.030
Meznerics, F. A., Fehérvári, P., Dembrovszky, F., Kovács, K. D., Kemény, L. V., Csupor, D., et al. (2022). Platelet-rich plasma in chronic wound management: a systematic review and meta-analysis of randomized clinical trials. Journal of Clinical Medicine, 11(24), 7532. https://doi.org/10.3390/jcm11247532
Salgado-Pacheco, V., Serra-Mas, M., & Otero-Viñas, M. (2024). Platelet-rich plasma therapy for chronic cutaneous wounds stratified by etiology: a systematic review of randomized clinical trials. Drugs & Therapy Perspectives, 40, 31–42. https://doi.org/10.1007/s40267-023-01044-7
OuYang, H., Tang, Y., Yang, F., et al. (2023). Platelet-rich plasma for the treatment of diabetic foot ulcer: A systematic review. Frontiers in Endocrinology, 14, 1256081. https://doi.org/10.3389/fendo.2023.1256081
Dai, J., Jiang, C., Sun, Y., et al. (2020). Autologous platelet-rich plasma treatment for patients with diabetic foot ulcers: A meta-analysis of randomized studies. Journal of Diabetes and Its Complications, 34(8), 107611. https://doi.org/10.1016/j.jdiacomp.2020.107611
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.